Lothar H Wieler

Publications of Lothar H Wieler

• tkt1, located on a novel pathogenicity island, is prevalent in avian and human extraintestinal pathogenic Escherichia coli.

  Authors: Ganwu Li, Subhashinie Kariyawasam, Kelly A Tivendale, Yvonne Wannemuehler, Christa Ewers, Lothar H Wieler, Catherine M Logue, Lisa K Nolan

  BMC microbiology. 04/2012; 12(1):51.

  ABSTRACT: BACKGROUND: Extraintestinal pathogenic Escherichia coli are important pathogens of human and animal hosts. Some human and avian extraintestinal pathogenic E. coli are indistinguishable onABSTRACT: BACKGROUND: Extraintestinal pathogenic Escherichia coli are important pathogens of human and animal hosts. Some human and avian extraintestinal pathogenic E. coli are indistinguishable on the basis of diseases caused, multilocus sequence and phylogenetic typing, carriage of large virulence plasmids and traits known to be associated with extraintestinal pathogenic E. coli virulence. RESULTS: The gene tkt1 identified by a previous signature-tagged transposon mutagenesis study, was found on a 16-kb genomic island of avian pathogenic Escherichia coli (APEC) O1, the first pathogenic Escherichia coli strain whose genome has been completely sequenced. tkt1 was present in 39.6% (38/96) of pathogenic Escherichia coli strains, while only 6.25% (3/48) of E. coli from the feces of apparently healthy chickens was positive. Further, tkt1 was predominantly present in extraintestinal pathogenic E. coli belonging to the B2 phylogenetic group, as compared to extraintestinal pathogenic E. coli of other phylogenetic groups. The tkt1-containing genomic island is inserted between the metE and ysgA genes of the E. coli K12 genome. Among different extraintestinal pathogenic E. coli of the B2 phylogenetic group, 61.7% of pathogenic Escherichia coli, 80.6% of human uropathogenic E.coli and 94.1% of human neonatal meningitis-causing E. coli, respectively, harbor a complete copy of this island; whereas, only a few avian fecal E. coli strains contained the complete island. Functional analysis showed that Tkt1 confers very little transketolase activity but is involved in peptide nitrogen metabolism. CONCLUSION: These results suggest tkt1 and its corresponding genomic island are frequently associated with avian and human ExPEC and are involved in bipeptide metabolism.

• Sharing More than Friendship - Nasal Colonization with Coagulase-Positive Staphylococci (CPS) and Co-Habitation Aspects of Dogs and Their Owners.

  Authors: Birgit Walther, Julia Hermes, Christiane Cuny, Lothar H Wieler, Szilvia Vincze, Yassmin Abou Elnaga, Ivonne Stamm, Peter A Kopp, Barbara Kohn, Wolfgang Witte, Andreas Jansen, Franz J Conraths, Torsten Semmler, Tim Eckmanns, Antina Lübke-Becker

  PloS one. 01/2012; 7(4):e35197.

  Since the relationship between dogs and their owners has changed, and dogs moved from being working dogs to family members in post-industrial countries, we hypothesized that zoonotic transmission ofSince the relationship between dogs and their owners has changed, and dogs moved from being working dogs to family members in post-industrial countries, we hypothesized that zoonotic transmission of opportunistic pathogens like coagulase positive staphylococci (CPS) is likely between dogs and their owners. CPS- nasal carriage, different aspects of human-to-dog relationship as well as potential interspecies transmission risk factors were investigated by offering nasal swabs and a questionnaire to dog owners (108) and their dogs (108) at a dog show in 2009. S. aureus was found in swabs of 20 (18.5%) humans and two dogs (1.8%), and spa types which correspond to well known human S. aureus lineages dominated (e.g. CC45, CC30 and CC22). Multilocus sequence typing (MLST) of the two canine strains revealed ST72 and ST2065 (single locus variant of ST34). Fifteen dogs (13.9%) and six owners (5.6%) harboured S. pseudintermedius, including one mecA-positive human isolate (MRSP). Pulsed field gel electrophoresis (PFGE) revealed that one dog/owner pair harboured indistinguishable S. pseudintermedius- isolates of ST33. Ten (48%) of the 21 S. pseudintermedius-isolates showed resistance towards more than one antimicrobial class. 88.9% of the dog owners reported to allow at least one dog into the house, 68.5% allow the dog(s) to rest on the sofa, 39.8% allow their dogs to come onto the bed, 93.5% let them lick their hands and 52.8% let them lick their face. Bivariate analysis of putative risk factors revealed that dog owners who keep more than two dogs have a significantly higher chance of being colonized with S. pseudintermedius than those who keep 1-2 dogs (p<0.05). In conclusion, CPS transmission between dog owners and their dogs is possible. Further investigation regarding interspecies transmission and the diverse adaptive pathways influencing the epidemiology of CPS (including MRSA and MRSP) in different hosts is needed.

• Methicillin-resistant staphylococci (MRS) and extended-spectrum beta-lactamases (ESBL)-producing Enterobacteriaceae in companion animals: nosocomial infections as one reason for the rising prevalence of these potential zoonotic pathogens in clinical samples.

  Authors: Lothar H Wieler, Christa Ewers, Sebastian Guenther, Birgit Walther, Antina Lübke-Becker

  International journal of medical microbiology : IJMM. 12/2011; 301(8):635-41.

  The ongoing change in the relationship between humans and companion animals is hallmarked by the increasing intensive care provided to companion animals in veterinary medicine, resulting in growingThe ongoing change in the relationship between humans and companion animals is hallmarked by the increasing intensive care provided to companion animals in veterinary medicine, resulting in growing numbers of high-risk animal patients. The emergence of nosocomial infections in small animal clinics is one of the major drawbacks of this development, especially in terms of multidrug-resistance and potentially zoonotic pathogens. This mini-review therefore addresses recent findings regarding the increasing prevalence of multi-resistant bacterial pathogens like methicillin-resistant staphylococci (MRS), including Staphylococcus aureus (MRSA) and Staphylococcus pseudintermedius (MRSP) as well as extended-spectrum beta-lactamases (ESBL)-producing Enterobacteriaceae in companion animals. Along with the steady increase of nosocomial infection rates in veterinary clinics, particular attention has recently been drawn to the genetic background of multi-resistant strains, resulting in the identification of certain genetic lineages which frequently appear in both, human and animal samples. These sequence types (ST), included ST254, ST8 and ST22 in terms of MRSA and ST131, ST405 and ST648 for ESBL-producing E. coli. The interspecies distribution of these STs resulted in the assumption that certain extended-host spectrum genotypes (EHSG) might exist both for MRS and ESBL-producing E. coli. These initial findings underline the necessity to investigate the major molecular or functional driving forces facilitating interspecies transferability of such EHSG strains. Due to the zoonotic potential of these multi-resistant bacteria, another aspect of the changing social role of companion animals needs to be addressed: the close contact of pets with their owners, resulting in presumptive new transmission and infection routes. We therefore envision retaliatory actions like initial surveillance and monitoring programs not only in livestock, but also particularly in companion animals. Interdisciplinary approaches including human and veterinary experts should be implemented to develop reliable investigation procedures with respect to the current reality of animal owners and their pets. Additionally, consequent basic hygienic measures, prudent use of antimicrobials in companion animals and efforts regarding implementation of antibiotic stewardships should be fostered.

• No evidence of the Shiga toxin-producing E. coli O104:H4 outbreak strain or enteroaggregative E. coli (EAEC) found in cattle faeces in northern Germany, the hotspot of the 2011 HUS outbreak area.

  Authors: Lothar H Wieler, Torsten Semmler, Inga Eichhorn, Esther M Antao, Bianca Kinnemann, Lutz Geue, Helge Karch, Sebastian Guenther, Astrid Bethe

  Gut pathogens. 11/2011; 3(1):17.

  ABSTRACT: Ruminants, in particular bovines, are the primary reservoir of Shiga toxin-producing E. coli (STEC), but whole genome analyses of the current German ESBL-producing O104:H4 outbreak strainABSTRACT: Ruminants, in particular bovines, are the primary reservoir of Shiga toxin-producing E. coli (STEC), but whole genome analyses of the current German ESBL-producing O104:H4 outbreak strain of sequence type (ST) 678 showed this strain to be highly similar to enteroaggregative E. coli (EAEC). Strains of the EAEC pathotype are basically adapted to the human host. To clarify whether in contrast to this paradigm, the O104:H4 outbreak strain and/or EAEC may also be able to colonize ruminants, we screened a total of 2.000 colonies from faecal samples of 100 cattle from 34 different farms - all located in the HUS outbreak region of Northern Germany - for genes associated with the O104:H4 HUS outbreak strain (stx2, terD, rfbO104, fliCH4), STEC (stx1, stx2, escV), EAEC (pAA, aggR, astA), and ESBL-production (blaCTX-M, blaTEM, blaSHV). The faecal samples contained neither the HUS outbreak strain nor any EAEC. As the current outbreak strain belongs to ST678 and displays an en-teroaggregative and ESBL-producing phenotype, we additionally screened selected strains for ST678 as well as the aggregative adhesion pattern in HEp-2 cells. However, we were unable to find any strains belonging to ST678 or showing an aggregative adhesion pattern. A high percentage of animals (28%) shed STEC, corroborating previous knowl-edge and thereby proving the validity of our study. One of the STEC also harboured the LEE pathogenicity island. In addition, eleven animals shed ESBL-producing E. coli. While we are aware of the limitations of our survey, our data support the theory, that, in contrast to other Shiga-toxin producing E. coli, cattle are not the reservoir for the O104:H4 outbreak strain or other EAEC, but that the outbreak strain seems to be adapted to humans or might have yet another reservoir, raising new questions about the epidemiology of STEC O104:H4.

• Adherent-invasive Escherichia coli phenotype displayed by intestinal pathogenic E. coli strains from cats, dogs, and swine.

  Authors: Margarita Martinez-Medina, Jesus Garcia-Gil, Nicolas Barnich, Lothar H Wieler, Christa Ewers

  Applied and environmental microbiology. 06/2011; 77(16):5813-7.

  The adherent-invasive Escherichia coli (AIEC) pathotype, which has been associated with Crohn's disease, shows similar traits to human and animal extraintestinal pathogenic E. coli (ExPEC) withThe adherent-invasive Escherichia coli (AIEC) pathotype, which has been associated with Crohn's disease, shows similar traits to human and animal extraintestinal pathogenic E. coli (ExPEC) with respect to their phylogenetic origin and virulence gene profiles. Here, we demonstrate that animal ExPEC strains generally do not share the AIEC phenotype. In contrast, this phenotype is very frequent among animal intestinal pathogenic E. coli (InPEC) strains, particularly of feline and canine origin, that genetically resemble ExPEC. These results strengthen the particular identity and disease specificity of the AIEC pathotype and the putative role animals might play in the transmission of AIEC-like strains to humans.

• E. coli Nissle 1917 Affects Salmonella adhesion to porcine intestinal epithelial cells.

  Authors: Peter Schierack, Sylvia Kleta, Karsten Tedin, Julius Tachu Babila, Sibylle Oswald, Tobias A Oelschlaeger, Rico Hiemann, Susanne Paetzold, Lothar H Wieler

  PloS one. 01/2011; 6(2):e14712.

  The probiotic Escherichia coli strain Nissle 1917 (EcN) has been shown to interfere in a human in vitro model with the invasion of several bacterial pathogens into epithelial cells, but theThe probiotic Escherichia coli strain Nissle 1917 (EcN) has been shown to interfere in a human in vitro model with the invasion of several bacterial pathogens into epithelial cells, but the underlying molecular mechanisms are not known. In this study, we investigated the inhibitory effects of EcN on Salmonella Typhimurium invasion of porcine intestinal epithelial cells, focusing on EcN effects on the various stages of Salmonella infection including intracellular and extracellular Salmonella growth rates, virulence gene regulation, and adhesion. We show that EcN affects the initial Salmonella invasion steps by modulating Salmonella virulence gene regulation and Salmonella SiiE-mediated adhesion, but not extra- and intracellular Salmonella growth. However, the inhibitory activity of EcN against Salmonella invasion always correlated with EcN adhesion capacities. EcN mutants defective in the expression of F1C fimbriae and flagellae were less adherent and less inhibitory toward Salmonella invasion. Another E. coli strain expressing F1C fimbriae was also adherent to IPEC-J2 cells, and was similarly inhibitory against Salmonella invasion like EcN. We propose that EcN affects Salmonella adhesion through secretory components. This mechanism appears to be common to many E. coli strains, with strong adherence being a prerequisite for an effective reduction of SiiE-mediated Salmonella adhesion.

• Virulence characteristics and genetic affinities of multiple drug resistant uropathogenic Escherichia coli from a semi urban locality in India.

  Authors: Savita Jadhav, Arif Hussain, Savita Devi, Ashutosh Kumar, Sana Parveen, Nageshwari Gandham, Lothar H Wieler, Christa Ewers, Niyaz Ahmed

  PloS one. 01/2011; 6(3):e18063.

  Extraintestinal pathogenic Escherichia coli (ExPEC) are of significant health concern. The emergence of drug resistant E. coli with high virulence potential is alarming. Lack of sufficient data onExtraintestinal pathogenic Escherichia coli (ExPEC) are of significant health concern. The emergence of drug resistant E. coli with high virulence potential is alarming. Lack of sufficient data on transmission dynamics, virulence spectrum and antimicrobial resistance of certain pathogens such as the uropathogenic E. coli (UPEC) from countries with high infection burden, such as India, hinders the infection control and management efforts. In this study, we extensively genotyped and phenotyped a collection of 150 UPEC obtained from patients belonging to a semi-urban, industrialized setting near Pune, India. The isolates representing different clinical categories were analyzed in comparison with 50 commensal E. coli isolates from India as well as 50 ExPEC strains from Germany. Virulent strains were identified based on hemolysis, haemagglutination, cell surface hydrophobicity, serum bactericidal activity as well as with the help of O serotyping. We generated antimicrobial resistance profiles for all the clinical isolates and carried out phylogenetic analysis based on repetitive extragenic palindromic (rep)-PCR. E. coli from urinary tract infection cases expressed higher percentages of type I (45%) and P fimbriae (40%) when compared to fecal isolates (25% and 8% respectively). Hemolytic group comprised of 60% of UPEC and only 2% of E. coli from feces. Additionally, we found that serum resistance and cell surface hydrophobicity were not significantly (p = 0.16/p = 0.51) associated with UPEC from clinical cases. Moreover, clinical isolates exhibited highest resistance against amoxicillin (67.3%) and least against nitrofurantoin (57.3%). We also observed that 31.3% of UPEC were extended-spectrum beta-lactamase (ESBL) producers belonging to serotype O25, of which four were also positive for O25b subgroup that is linked to B2-O25b-ST131-CTX-M-15 virulent/multiresistant type. Furthermore, isolates from India and Germany (as well as global sources) were found to be genetically distinct with no evidence to espouse expansion of E. coli from India to the west or vice-versa.

• Extended-Spectrum Beta-Lactamases Producing E. coli in Wildlife, yet Another Form of Environmental Pollution?

  Authors: Sebastian Guenther, Christa Ewers, Lothar H Wieler

  Frontiers in microbiology. 01/2011; 2:246.

  Wildlife is normally not exposed to clinically used antimicrobial agents but can acquire antimicrobial resistant bacteria through contact with humans, domesticated animals and the environment, whereWildlife is normally not exposed to clinically used antimicrobial agents but can acquire antimicrobial resistant bacteria through contact with humans, domesticated animals and the environment, where water polluted with feces seems to be the most important vector. Escherichia coli, an ubiquitous commensal bacterial species colonizing the intestinal tract of mammals and birds, is also found in the environment. Extended-spectrum beta-lactamases producing E. coli (ESBL-E. coli) represent a major problem in human and veterinary medicine, particular in nosocomial infections. Additionally an onset of community-acquired ESBL-E. coli infections and an emergence in livestock farming has been observed in recent years, suggesting a successful transmission as well as persistence of ESBL-E. coli strains outside clinical settings. Another parallel worldwide phenomenon is the spread of ESBL-E. coli into the environment beyond human and domesticated animal populations, and this seems to be directly influenced by antibiotic practice. This might be a collateral consequence of the community-onset of ESBL-E. coli infections but can result (a) in a subsequent colonization of wild animal populations which can turn into an infectious source or even a reservoir of ESBL-E. coli, (b) in a contribution of wildlife to the spread and transmission of ESBL-E. coli into fragile environmental niches, (c) in new putative infection cycles between wildlife, domesticated animals and humans, and (d) in problems in the medical treatment of wildlife. This review aims to summarize the current knowledge on ESBL-E. coli in wildlife, in turn underlining the need for more large scale investigations, in particular sentinel studies to monitor the impact of multiresistant bacteria on wildlife.

• Companion animals: a relevant source of extended-spectrum β-lactamase-producing fluoroquinolone-resistant Citrobacter freundii.

  Authors: Christa Ewers, Astrid Bethe, Lothar H Wieler, Sebastian Guenther, Ivonne Stamm, Peter A Kopp, Mirjam Grobbel

  International journal of antimicrobial agents. 11/2010; 37(1):86-7.

• Antimicrobial susceptibilities and occurrence of resistance genes in bovine Helcococcus ovis isolates.

  Authors: Sabine Bilk, Marcel Nordhoff, Christoph Schulze, Lothar H Wieler, Peter Kutzer

  Veterinary microbiology. 11/2010; 149(3-4):488-91.

  The aim of the study was to investigate antimicrobial susceptibilities of bovine Helcococcus ovis isolates and to detect genes encoding for H. ovis erythromycin and tetracycline resistance.The aim of the study was to investigate antimicrobial susceptibilities of bovine Helcococcus ovis isolates and to detect genes encoding for H. ovis erythromycin and tetracycline resistance. Twenty-nine isolates were included and the minimal inhibitory concentrations (MICs) of seven antimicrobials were determined using test conditions as given in an approved CLSI guideline for the pyridoxal-dependent Abiotrophia spp. and Granulicatella spp. Furthermore, the macrolide resistance phenotype was examined by the erythromycin-clindamycin double-disk test (D-zone test). Erythromycin MICs of ≥ 8 μg/ml were found in three (10%) isolates which also presented the macrolide, lincosamide, and streptogramin B (MLS(B)) resistance phenotype, either constitutive or inducible. The erm(B) gene was detected in one of these isolates. Increased tetracycline MICs (≥ 8 μg/ml) were obtained for 24 (83%) isolates, mostly associated with the tet(M) gene alone (n=21) or both the tet(L) and tet(M) genes (n=2). The MICs determined for penicillin, ampicillin, amoxicillin-clavulanic acid, and cephalothin did not indicate resistance to these antimicrobials. The study suggests that resistance to MLS(B) antimicrobials and tetracycline is frequent in H. ovis. Moreover, this is the first report about occurrence of the resistance genes erm(B), tet(L), and tet(M) in the Helcococcus genus.

• First insights into antimicrobial resistance among faecal Escherichia coli isolates from small wild mammals in rural areas.

  Authors: Sebastian Guenther, Mirjam Grobbel, Katrin Heidemanns, Mathias Schlegel, Rainer G Ulrich, Christa Ewers, Lothar H Wieler

  The Science of the total environment. 08/2010; 408(17):3519-22.

  Wild rodents can be carriers of antimicrobial resistant Escherichia coli. As rodents are known to be involved in the transmission of bacteria of human and animal health concern, they could likewiseWild rodents can be carriers of antimicrobial resistant Escherichia coli. As rodents are known to be involved in the transmission of bacteria of human and animal health concern, they could likewise contribute to the dissemination of antimicrobial resistant bacteria in the environment. The aim of this study was therefore to get first insights into the antimicrobial resistance status among E. coli isolated from wild small mammals in rural areas. We tested 188 faecal isolates from eight rodent and one shrew species originating from Germany. Preselected resistant isolates were screened by minimal inhibitory concentration (MIC) testing or agar diffusion test and subsequent PCR analysis of resistance genes. The prevalence of antimicrobial resistant isolates was low with only 5.5% of the isolates exhibiting resistant phenotypes against at least one antimicrobial compound including beta-lactams, tetracyclines, aminoglycosides and sulfonamides. These results suggest a minor role of wild rodents from rural areas in the cycle of transmission and spread of antimicrobial resistant E. coli into the environment. Nevertheless E. coli with multiple antimicrobial resistances were significantly more often detected in wildlife rodents originating from areas with high livestock density suggesting a possible transmission from livestock to wild rodents.

• Antimicrobial resistance profiles of Escherichia coli from common European wild bird species.

  Authors: Sebastian Guenther, Mirjam Grobbel, Antina Lübke-Becker, Andreas Goedecke, Nicole D Friedrich, Lothar H Wieler, Christa Ewers

  Veterinary microbiology. 07/2010; 144(1-2):219-25.

  The emergence and spread of multiresistant bacteria in natural environments constitute a serious impact on animal and human health. To gain more insight into the role of wild birds as carriers andThe emergence and spread of multiresistant bacteria in natural environments constitute a serious impact on animal and human health. To gain more insight into the role of wild birds as carriers and reservoir of multiresistant Escherichia coli we tested a broad spectrum of common European bird species for the occurrence of E. coli strains and their antimicrobial resistance by minimal inhibitory concentration testing and PCR analysis of several resistance genes. Nine of the 187 E. coli isolates (4.8%) exhibited multiresistant phenotypes including resistances against beta-lactams, aminoglycosides, fluoroquinolones, tetracyclines and sulfonamides. By comparing avian E. coli resistance frequencies with frequencies known for E. coli isolated from livestock and companion animals analogous profiles were identified. Multiresistant E. coli strains were isolated from synanthropic avian species as well as from birds of prey, waterfowl and passerines. By that, all these avian hosts are suggested to represent a considerable reservoir of resistant E. coli strains. Consequently wild birds might constitute a potential hazard to human and animal health by transmitting multiresistant strains to waterways and other environmental sources via their faecal deposits.

• Identification of protective and broadly conserved vaccine antigens from the genome of extraintestinal pathogenic Escherichia coli.

  Authors: Danilo Gomes Moriel, Isabella Bertoldi, Angela Spagnuolo, Sara Marchi, Roberto Rosini, Barbara Nesta, Ilaria Pastorello, Vanja A Mariani Corea, Giulia Torricelli, Elena Cartocci [......] Luke J Tallon, Steven Sullivan, Lothar H Wieler, Christa Ewers, Derek Pickard, Gordon Dougan, Maria Rita Fontana, Rino Rappuoli, Mariagrazia Pizza, Laura Serino

  Proceedings of the National Academy of Sciences of the United States of America. 05/2010; 107(20):9072-7.

  Extraintestinal pathogenic Escherichia coli (ExPEC) are a common cause of disease in both mammals and birds. A vaccine to prevent such infections would be desirable given the increasing antibioticExtraintestinal pathogenic Escherichia coli (ExPEC) are a common cause of disease in both mammals and birds. A vaccine to prevent such infections would be desirable given the increasing antibiotic resistance of these bacteria. We have determined the genome sequence of ExPEC IHE3034 (ST95) isolated from a case of neonatal meningitis and compared this to available genome sequences of other ExPEC strains and a few nonpathogenic E. coli. We found 19 genomic islands present in the genome of IHE3034, which are absent in the nonpathogenic E. coli isolates. By using subtractive reverse vaccinology we identified 230 antigens present in ExPEC but absent (or present with low similarity) in nonpathogenic strains. Nine antigens were protective in a mouse challenge model. Some of them were also present in other pathogenic non-ExPEC strains, suggesting that a broadly protective E. coli vaccine may be possible. The gene encoding the most protective antigen was detected in most of the E. coli isolates, highly conserved in sequence and found to be exported by a type II secretion system which seems to be nonfunctional in nonpathogenic strains.

• Emergence of human pandemic O25:H4-ST131 CTX-M-15 extended-spectrum-beta-lactamase-producing Escherichia coli among companion animals.

  Authors: Christa Ewers, Mirjam Grobbel, Ivonne Stamm, Peter A Kopp, Ines Diehl, Torsten Semmler, Angelika Fruth, Janine Beutlich, Beatriz Guerra, Lothar H Wieler, Sebastian Guenther

  The Journal of antimicrobial chemotherapy. 04/2010; 65(4):651-60.

  In view of the intercontinental emergence of Escherichia coli clone O25:H4-ST131 producing CTX-M-15 extended-spectrum beta-lactamase (ESBL) in human clinical settings it would be of great interest toIn view of the intercontinental emergence of Escherichia coli clone O25:H4-ST131 producing CTX-M-15 extended-spectrum beta-lactamase (ESBL) in human clinical settings it would be of great interest to explore its existence in animals to unravel a possible reservoir function and the origin and transmission of this group of multiresistant strains. A total of 177 clinical phenotypically ESBL-producing E. coli isolates, mainly obtained from companion animals with urinary tract infections, wound infections and diarrhoea, were collected in a veterinary diagnostic laboratory covering a European-wide service area. They were screened for molecular subtype O25b and multilocus sequence type 131. O25b-ST131 isolates were subsequently tested for ESBL types, and phenotypic and genotypic resistance determinants. Further characterization of the strains was performed by PFGE and virulence gene typing. Ten (5.6%) of 177 phenotypically ESBL-producing E. coli isolates, nine strains from dogs and one strain from a horse, were allocated to the B2-O25b-ST131 lineage. Nine of these isolates harboured a CTX-M-15-type beta-lactamase enzyme while one strain possessed an SHV-12-type ESBL. Macrorestriction analysis revealed a cluster formation of six of the animal CTX-M-15-type ESBL-producing strains from five different European countries together with a human control strain constituting a group of clonally related strains at a similarity value of 87.0%. Our findings demonstrate that the group of clonally related human B2-O25:H4-ST131 CTX-M-15-type ESBL-producing E. coli strains is present in companion animals from various European countries. This highlights the possibility of inter-species transmission of these multiresistant strains from human to animal and vice versa.

• Detection of pandemic B2-O25-ST131 Escherichia coli harbouring the CTX-M-9 extended-spectrum beta-lactamase type in a feral urban brown rat (Rattus norvegicus).

  Authors: Sebastian Guenther, Mirjam Grobbel, Janine Beutlich, Beatriz Guerra, Rainer G Ulrich, Lothar H Wieler, Christa Ewers

  The Journal of antimicrobial chemotherapy. 03/2010; 65(3):582-4.

• CTX –M 15 type extended spectrum beta lactamases producing E. coli from wild birds in Germany

  Authors: Sebastian Guenther, Mirjam Grobbel, Janine Beutlich, Astrid Bethe, Antina Lübke-Becker, Andreas Goedecke, Nicole D. Friedrich, Beatriz Guerra-Roman, Lothar H. Wieler, Christa Ewers 2010

  Environmental Microbiology Reports. 01/2010; 2:641–645.

• Widespread rapid emergence of a distinct methicillin- and multidrug-resistant Staphylococcus pseudintermedius (MRSP) genetic lineage in Europe.

  Authors: Claudia Ruscher, Antina Lübke-Becker, Torsten Semmler, Claus-G Wleklinski, Angela Paasch, Alexandra Soba, Ivonne Stamm, Peter Kopp, Lothar H Wieler, Birgit Walther

  Veterinary microbiology. 01/2010; 144(3-4):340-6.

  In order to gain a deeper insight into the phylogenetic background and diversity of methicillin-resistant S. pseudintermedius (MRSP) of animal origin, genetic relationships and clonal distributionIn order to gain a deeper insight into the phylogenetic background and diversity of methicillin-resistant S. pseudintermedius (MRSP) of animal origin, genetic relationships and clonal distribution among 146 European MRSP were examined using different molecular and phenotypical typing approaches. MRSP strains were derived from clinical microbiological specimens (mainly of small animal origin) sent in for diagnostic purposes from various veterinary facilities between 2005 and 2008. Pulsed-field gel electrophoresis (PFGE) of SmaI-macrorestriction fragments allowed differentiation of five PFGE-clusters that were subdivided into further distinct subtypes. Representatives of each PFGE subtype were analyzed by multilocus sequence typing (MLST) for assignment of sequence types (ST). With one exception (ST5), all these MRSP strains belonged to ST71. Furthermore, assessment of spa-typing results revealed that the majority of all strains harboured spa type t02. Further sporadically detected spa types t05 and t06 as well as two new types (t15 and t23), were found to be closely related to t02. According to PCR-based SCCmec-typing, SCCmecIII was the most prevalent type (n=138), and solely one non-typeable variant was identified in several strains (n=8). In addition, all strains were tested positive by PCR for the leukotoxin encoding operon LukI and the Staphylococcus intermedius-exfoliative toxin (SIET), respectively. Our cumulative data indicate a recent emergence of a certain multidrug-resistant MRSP-lineage (ST71) in central and southern European countries during the last few years.

• The GimA locus of extraintestinal pathogenic E. coli: does reductive evolution correlate with habitat and pathotype?

  Authors: Timo Homeier, Torsten Semmler, Lothar H Wieler, Christa Ewers

  PloS one. 01/2010; 5(5):e10877.

  IbeA (invasion of brain endothelium), which is located on a genomic island termed GimA, is involved in the pathogenesis of several extraintestinal pathogenic E. coli (ExPEC) pathotypes, includingIbeA (invasion of brain endothelium), which is located on a genomic island termed GimA, is involved in the pathogenesis of several extraintestinal pathogenic E. coli (ExPEC) pathotypes, including newborn meningitic E. coli (NMEC) and avian pathogenic E. coli (APEC). To unravel the phylogeny of GimA and to investigate its island character, the putative insertion locus of GimA was determined via Long Range PCR and DNA-DNA hybridization in 410 E. coli isolates, including APEC, NMEC, uropathogenic (UPEC), septicemia-associated E. coli (SEPEC), and human and animal fecal isolates as well as in 72 strains of the E. coli reference (ECOR) collection. In addition to a complete GimA (approximately 20.3 kb) and a locus lacking GimA we found a third pattern containing a 342 bp remnant of GimA in this strain collection. The presence of GimA was almost exclusively detected in strains belonging to phylogenetic group B2. In addition, the complete GimA was significantly more frequent in APEC and NMEC strains while the GimA remnant showed a higher association with UPEC strains. A detailed analysis of the ibeA sequences revealed the phylogeny of this gene to be consistent with that obtained by Multi Locus Sequence Typing of the strains. Although common criteria for genomic islands are partially fulfilled, GimA rather seems to be an ancestral part of phylogenetic group B2, and it would therefore be more appropriate to term this genomic region GimA locus instead of genomic island. The existence of two other patterns reflects a genomic rearrangement in a reductive evolution-like manner.

• Shiga toxin producing Escherichia coli: identification of non-O157:H7-Super-Shedding cows and related risk factors.

  Authors: Andrea Menrath, Lothar H Wieler, Katrin Heidemanns, Torsten Semmler, Angelika Fruth, Nicole Kemper

  Gut pathogens. 01/2010; 2(1):7.

  Shiga toxin producing Escherichia coli (STEC) are an important cause of human gastro-enteritis and extraintestinal sequelae, with ruminants, especially cattle, as the major source of infection andShiga toxin producing Escherichia coli (STEC) are an important cause of human gastro-enteritis and extraintestinal sequelae, with ruminants, especially cattle, as the major source of infection and reservoir. In this study, the fecal STEC shedding of 133 dairy cows was analyzed over a period of twelve months by monthly sampling with the aim to investigate shedding patterns and risk factors. Overall, 24.7% (in total 407) of 1,646 fecal samples were tested positive for stx by PCR with inner-herd prevalences on the different farms of 11.1% to 32.3%. At individual levels, cows were stx-positive on zero to eight consecutive samplings. According to a strictly longitudinal definition of Super-Shedding, in the present study 14 cows were identified as Super-Shedders of non-O157 serotypes.Significant risk factors for the shedding of STEC were the month of sampling, the number of lactations and days in lactation, the nutritional condition, the somatic cell count and the content of protein in milk. Most notably, the presence of STEC Super-Shedding cows in the herd was a significant risk factor, revealing that STEC Super-Shedding is not restricted to STEC O157:H7 alone. These data have implications for possible interventions, as removing single non-O157:H7 STEC Super-Shedding cattle from farms would significantly reduce STEC burden.

• Detection of bla(CTX-M-15) extended-spectrum beta-lactamase genes in Escherichia coli from hospital patients in Nigeria.

  Authors: Olowe Adekunle, Mirjam Grobbel, Britta Büchter, Antina Lübke-Becker, A Fruth, Lothar H Wieler

  International journal of antimicrobial agents. 12/2009;